Method of administering and using VEGF inhibitors for the treatment of human cancer

ABSTRACT

A method of treating a human patient suffering from cancer, comprising administering an effective amount of a vascular endothelial growth factor (VEGF) trap antagonist to the human patient, the method comprising: (a) administering to the patient an initial dose of at least approximately 0.3 mg/kg of the VEGF antagonist; and (b) administering to the patient a plurality of subsequent doses of the VEGF antagonist in an amount that is approximately the same or less of the initial dose, wherein the subsequent doses are separated in time from each other by at least one day. The methods of the invention are useful for treating a human cancer selected from the group consisting of renal cell carcinoma, pancreatic carcinoma, breast cancer, prostate cancer, colorectal cancer, malignant mesothelioma, multiple myeloma, ovarian cancer, and melanoma. The invention is further useful for treating a condition which benefits from the reduction of VEGFA and placental growth factor (PLGF).

RELATED APPLICATIONS

This application is a divisional application of U.S. application Ser.No. 11/149,738 filed 10 Jun. 2005, now allowed, which claims the benefitunder 35 USC 119(e) to U.S. Ser. No. 60/578,499 filed 10 Jun. 2004,which applications are incorporated by reference in their entirety.

FIELD OF THE INVENTION

The invention relates to methods of promoting regression of tumors andmetastases by inhibiting vascular endothelial growth factor (VEGF)activity.

DESCRIPTION OF RELATED ART

Vascular endothelial growth factor (VEGF) expression is nearlyubiquitous in human cancer, consistent with its role as a key mediatorof tumor neoangiogenesis. Blockade of VEGF function, by binding to themolecule or its VEGFR-2 receptor, inhibits growth of implanted tumorcells in multiple different xenograft models (see, for example, Gerberet al. (2000) Cancer Res. 60:6253-6258). A soluble VEGF fusion proteinantagonist, termed a “VEGF_(R1R2) trap” or “VEGF trap” antagonist hasbeen described (Kim et al. (2002) Proc. Natl. Acad. Sci. USA99:11399-404; Holash et al. (2002) Proc. Natl. Acad. Sci. USA99:11393-8), which applications are specifically incorporated byreference in their entirety.

BRIEF SUMMARY OF THE INVENTION

In a first aspect, the invention features a method of treating a humanpatient suffering from cancer, comprising administering an effectiveamount of a vascular endothelial growth factor (VEGF) fusion proteintrap antagonist to the human patient, the method comprising: (a)administering to the patient an initial dose of at least approximately0.3 mg/kg of the VEGF trap antagonist; and (b) administering to thepatient a plurality of subsequent doses of the VEGF trap antagonist inan amount that is approximately the same or less of the initial dose,wherein the subsequent doses are separated in time from each other by atleast one day. The dosing regimen of the invention allows earlyattainment of an efficacious target trough serum concentration byproviding an initial dose or doses of VEGF trap antagonist followed bysubsequent doses of equal or smaller amounts of trap (greater frontloading). The efficacious target trough serum concentration is reachedin 4 weeks or less, preferably 3 weeks or less, more preferably 2 weeksor less, and most preferably 1 week or less, including 1 day or less.The target serum concentration is thereafter maintained by theadministration of maintenance doses of equal or smaller amounts for theremainder of the treatment regimen or until suppression of diseasesymptoms is achieved.

In specific embodiments, the initial dose of the VEGF fusion proteinantagonist is in the range of approximately between 0.3 mg per kg bodyweight (mg/kg/kg) to 30 mg/kg. In a more specific embodiment, theinitial dose is in the range of approximately 0.5 mg/kg to 10 mg/kg. Inan even more specific embodiment, the initial dose is in the range ofapproximately 1 mg/kg to 6 mg/kg. Preferably, the cumulative weekly doseis in the range of 0.3 to 30 mg/kg.

In specific embodiments, at least one subsequent dose of the VEGF fusionprotein antagonist is in the range of approximately between 0.3 mg/kgbody weight to 30 mg/kg. In a more specific embodiment, at least onesubsequent dose is in the range of approximately 0.5 mg/kg to 10 mg/kg.In an even more specific embodiment, at least one; subsequent dose is inthe range of approximately 1 mg/kg to 6 mg/kg.

In one embodiment, subsequent doses are separated in time from eachother by at least 1 day, at least 1 week, at least 2 weeks, at least 3weeks, at least 1 month, at least 2 months, or at least 3 months.According to the invention, the cycle of dosing is preferably repeatedas necessary to achieve suppression of the disease symptoms.

The method of the invention may be used to treat primary and/ormetastatic tumors arising in the brain and meninges, oropharynx, lungand bronchial tree, gastrointestinal tract, male and female reproductivetract, muscle, bone, skin and appendages, connective tissue, spleen,immune system, blood forming cells and bone marrow, liver and urinarytract, and special sensory organs such as the eye. More specifically,the human patient treated by the method of the invention is a patientdiagnosed with one of the following cancers: renal cell carcinoma,pancreatic carcinoma, breast cancer, prostate cancer, colorectal cancer,malignant mesothelioma, multiple myeloma, ovarian cancer, or melanoma.In a specific embodiment, the cancer being treated is renal cellcarcinoma. In another embodiment, the cancer being treated is pancreaticcarcinoma. In another embodiment, the cancer being treated is breastcancer. In another embodiment, the cancer being treated is colorectalcancer. In another embodiment, the cancer being treated is malignantmesothelioma. In another embodiment, the cancer being treated ismultiple myeloma. In another embodiment, the cancer being treated isovarian cancer. In another embodiment, the cancer being treated ismelanoma. In another embodiment, the cancer being treated is non-smallcell lung cancer. In another embodiment, the cancer being treated isprostate cancer.

The VEGF fusion protein trap antagonist is a dimer comprising two fusionproteins each composed of immunoglobulin (Ig)—like domains from twodifferent VEGF receptors fused to a multimerizing component, whereineach fusion protein is capable of forming a higher order complex throughinteraction of multimerizing components on different fusion proteins.The VEGF trap antagonist useful in the method of the present inventionis a dimer capable of binding both vascular endothelial growth factor A(VEGFA) and placental growth factor (PLGF), and is selected from thegroup consisting of acetylated Flt-1(1-3)-Fc, Flt-1 (1-3_(R->N))-Fc,FLt-1 (1-3_(ΔB))-Fc, Flt-1(2-3_(ΔB))-Fc, Flt-1(2-3)-Fc,Flt-1D2-VEGFR3D3-FcΔC1(a), Flt-1D2-Flk-1D3-FcΔC1(a), andVEGFR1R2-FcΔC1(a). In a specific and preferred embodiment, the VEGF trapantagonist is VEGFR1R2-FcΔC1(a) (also termed VEGF trap_(R1R2)) havingthe nucleotide sequence set forth in SEQ ID NO: 1 and the amino acidsequence set forth in SEQ ID NO: 2. The invention encompasses the use ofa VEGF trap that is at least 90%, 95%, 98%, or at least 99% homologouswith the nucleotide sequence set forth in SEQ ID NO: 1 and/or the aminoacid sequence set forth in SEQ ID NO:2.

Administration of the agent may be by any method known in the art,including subcutaneous, intramuscular, intradermal, intraperitoneal,intravenous, intranasal, or oral routes of administration. In apreferred embodiment, the initial dose is administered by subcutaneousinjection or intravenous injection. In further embodiments, thesubsequent doses are administered by subcutaneous injection. In apreferred embodiment, the initial dose and at least one subsequent doseare administered by subcutaneous injection.

In a second aspect, the invention features a method of treating a humanpatient susceptible to or diagnosed with a disorder which is inhibitedby an agent capable of blocking or inhibiting vascular endothelialgrowth factor A (VEGF A), wherein the agent capable of blocking orinhibiting VEGFA is a VEGF trap antagonist, the method comprising: (a)administering to the patient an initial dose of at least approximately0.3 mg/kg of the VEGF trap; and (b) administering to the patient aplurality of subsequent doses of the VEGF trap in an amount that isapproximately the same or less of the initial dose, wherein thesubsequent doses are separated in time from each other by at least oneday. In a specific and preferred embodiment, the VEGF trap antagonist isVEGFR1R2-FcΔC1 (a) (also termed VEGF trap_(R1R2)) having the nucleotidesequence set forth in SEQ ID NO: 1 and the amino acid sequence set forthin SEQ ID NO: 2.

In a third embodiment, the invention features a therapeutic method ofthe invention optionally combined with a second chemotherapeutic agent.Chemotherapeutic agents combinable with administration of VEGF trapinclude, for example, anti-VEGF antibodies, anthracycline derivatives,such as doxorubicin or epirubicin taxol, and taxoid derivatives such aspaclitaxel (Taxol®) and related derivatives.

Other objects and advantages will become apparent from a review of theensuing detailed description.

DETAILED DESCRIPTION

Before the present methods are described, it is to be understood thatthis invention is not limited to particular methods, and experimentalconditions described, as such methods and conditions may vary. It isalso to be understood that the terminology used herein is for thepurpose of describing particular embodiments only, and is not intendedto be limiting, since the scope of the present invention will be limitedonly the appended claims.

As used in this specification and the appended claims, the singularforms “a”, “an”, and “the” include plural references unless the contextclearly dictates otherwise. Thus for example, a reference to “a method”includes one or more methods, and/or steps of the type described hereinand/or which will become apparent to those persons skilled in the artupon reading this disclosure and so forth.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although any methods andmaterials similar or equivalent to those described herein can be used inthe practice or testing of the present invention, the preferred methodsand materials are now described. All publications mentioned herein areincorporated herein by reference in their entirety.

GENERAL DESCRIPTION

Vascular endothelial growth factor/vascular permeability factor (VEGF)was initially identified as a tumor-derived factor capable of increasingvascular permeability. It was subsequently found to be a proliferativefactor for endothelial cells. In the embryo, VEGF is absolutelyessential for the development of the vasculature. In the adult, VEGF isup-regulated in a variety of normal and pathological processesassociated with increased vascular permeability and angiogenesis.

The family of VEGF-related angiogenic growth factors is comprised ofVEGF itself (VEGF-A) and the related proteins VEGF-B, -C, -D and E, andplacental growth factor (PLGF). In addition, there are at least fourdifferent isoforms of VEGF-A. However, as some members of the familyhave only recently been identified, their biological importance is stillpoorly understood. The actions of VEGF and its related factors aremediated by a group of three receptor tyrosine kinases, VEGFR1, VEGFR2and VEGFR3.

The importance of VEGF in tumor angiogenesis has been demonstrated in anumber of animal models, where blocking VEGF signaling by a variety ofstrategies has proven effective at decreasing angiogenesis andinhibiting tumor growth (Gourley and Williamson (2000) Curr. Pharm. Des.6:417-39). The permeability-inducing properties of VEGF are also ofpathological importance, for example in edema formation, ascites andpleural effusions related to cancer. The degree of vascularization andof VEGF production have been proposed as prognostic factors for manytypes of solid and hematological malignancies (reviewed by Poon et al(2001) J. Clin. Oncol. 19:1207-1225).

Consistent with predictions from animal studies, blockade of VEGF usinga humanized monoclonal antibody has emerged reporting promising resultsin cancer patients, based on preliminary reports from early clinicaltrials (Bergsland et al. (2000) ASCO Abstract #939). The VEGF fusionprotein trap antagonist, because of its greater affinity for VEGF andits ability to bind other VEGF family members such as the PIGFs, is apotent and useful anti-cancer therapeutic agent.

DEFINITIONS

By the term “therapeutically effective dose” is meant a dose thatproduces the desired effect for which it is administered. The exact dosewill depend on the purpose of the treatment, and will be ascertainableby one skilled in the art using known techniques (see, for example,Lloyd (1999) The Art, Science and Technology of PharmaceuticalCompounding). Efficacy can be measured in conventional ways, dependingon the condition to be treated. For cancer therapy, efficacy can, forexample, be measured by assessing the time to disease progression (TTP),or determining the response rates (RR). Therapeutically effective amountalso refers to a target serum concentration, such as a trough serumconcentration, that has been shown to be effective in suppressingdisease symptoms when maintained for a period of time.

The terms “cancer” and “cancerous” refer to or describe thephysiological condition in mammals that is typically characterized byunregulated cell growth. Examples of cancer include, but are not limitedto, carcinoma, lymphoma, blastoma, sarcoma, and leukemia. Moreparticular examples of such cancers include squamous cell cancer,small-cell lung cancer, non-small cell lung cancer, gastrointestinalcancer, pancreatic cancer, glioblastoma, cervical cancer, ovariancancer, liver cancer, bladder cancer, hepatoma, breast cancer, prostatecancer, colon cancer, colorectal cancer, endometrial carcinoma, salivarygland carcinoma, kidney cancer, prostate cancer, vulval cancer, thyroidcancer, hepatic carcinoma and various types of head and neck cancer.More specifically, the methods of the invention are useful for treatingany condition or disease which is ameliorated or inhibited with a VEGFinhibitor. Accordingly, when the disease or condition is cancer, thecancer treated by the method of the invention is one which isameliorated or inhibited by administration of a VEGF inhibitor.

By the term “blocker”, “inhibitor”, or “antagonist” is meant a substancethat retards or prevents a chemical or physiological reaction orresponse. Common blockers or inhibitors include but are not limited toantisense molecules, antibodies, antagonists and their derivatives. Morespecifically, an example of a VEGF blocker or inhibitor is a VEGFreceptor-based antagonist including, for example, an anti-VEGF antibody,or a VEGF trap antagonist such as VEGFR1R2-FcΔC1(a) (SEQ ID NOs:1-2).For a complete description of VEGF-receptor based antagonists includingVEGFR1R2-FcΔC1(a), see PCT publication WO/00/75319, the contents ofwhich is herein incorporated by reference in its entirety.

The term “package insert” is used to refer to instructions customarilyincluded in commercial packages of therapeutic products, that containinformation about the indications, usage, dosage, administration,contraindications and/or warnings concerning the use of such therapeuticproducts.

The term “serum concentration,” “serum drug concentration,” or “serumVEGF trap concentration” refers to the concentration of a drug, such asthe VEGF fusion protein trap antagonist, in the blood serum or plasma ofan animal or human patient being treated with the drug. Serumconcentration is preferably determined by immunoassay. Preferably, theimmunoassay is an ELISA according to the procedure disclosed herein.

The term “peak serum concentration” refers to the maximal serum drugconcentration shortly after delivery of the drug into the animal orhuman patient, after the drug has been distributed throughout the bloodsystem, but before significant tissue distribution, metabolism orexcretion of drug by the body has occurred.

The term “trough serum concentration” refers to the serum drugconcentration at a time after delivery of a previous dose andimmediately prior to delivery of the next subsequent dose of drug in aseries of doses. Generally, the trough serum concentration is a minimumsustained efficacious drug concentration in the series of drugadministrations. Also, the trough serum concentration is frequentlytargeted as a minimum serum concentration for efficacy because itrepresents the serum concentration at which another dose of drug is tobe administered as part of the treatment regimen. If the delivery ofdrug is by intravenous administration, the trough serum concentration ismost preferably attained within 1 day of a front loading initial drugdelivery. If the delivery of drug is by subcutaneous administration, thepeak serum concentration is preferably attained in 3 days or less.According to the invention, the trough serum concentration is preferablyattained in 4 weeks or less, preferably 3 weeks or less, more preferably2 weeks or less, most preferably in 1 week or less, including 1 day orless using any of the drug delivery methods disclosed herein.

The term “intravenous infusion” refers to introduction of a drug intothe vein of an animal or human patient over a period of time greaterthan approximately 5 minutes, preferably between approximately 30 to 90minutes, although, according to the invention, intravenous infusion isalternatively administered for 10 hours or less.

The term “subcutaneous administration” refers to introduction of a drugunder the skin of an animal or human patient, preferable within a pocketbetween the skin and underlying tissue, by relatively slow, sustaineddelivery from a drug receptacle. The pocket may be created by pinchingor drawing the skin up and away from underlying tissue.

The term “front loading” when referring to drug administration is meantto describe an initially higher dose followed by the same or lower dosesat intervals. The initial higher dose or doses are meant to more rapidlyincrease the animal or human patient's serum drug concentration to anefficacious target serum concentration. According to the presentinvention, front loading is achieved by an initial dose or dosesdelivered over three weeks or less that causes the animal's or patient'sserum concentration to reach a target serum trough concentration.Preferably, the initial front loading dose or series of doses isadministered in two weeks or less, more preferably in 1 week or less,including 1 day or less. Most preferably, where the initial dose is asingle dose and is not followed by a subsequent maintenance dose for atleast 1 week, the initial dose is administered in 1 day or less. Wherethe initial dose is a series of doses, each dose is separated by atleast 3 hours, but not more than 3 weeks or less, preferably 2 weeks orless, more preferably 1 week or less, most preferably 1 day or less.

The VEGF Fusion Protein Trap Antagonist

In a preferred embodiment, the VEGF trap is a receptor-Fc fusion proteinconsisting of the principal ligand-binding portions of the human VEGFR1and VEGFR2 receptor extracellular domains fused to the Fc portion ofhuman IgG1. Specifically, the VEGF Trap consists essentially of Igdomain 2 from VEGFR1, which is fused to Ig domain 3 from VEGFR2, whichin turn is fused to the Fc domain of IgG1 (SEQ ID NO:2).

In a preferred embodiment, an expression plasmid encoding the VEGF trapis transfected into CHO cells, which secrete VEGF trap into the culturemedium. The resulting VEGF trap is a dimeric glycoprotein with a proteinmolecular weight of 97 kDa and contains ˜15% glycosylation to give atotal molecular weight of 115 kDa.

Since the VEGF trap binds its ligands using the binding domains ofhigh-affinity receptors, it has a greater affinity for VEGF than domonoclonal antibodies. The VEGF trap binds VEGF-A (K_(D)=0.5 μM), PLGF1(K_(D)=1.3 nM), and PLGF2 (K_(D)=50 pM); binding to other VEGF familymembers has not yet been fully characterized.

Treatment Population

The method of the invention may be used to treat tumors arising in thebrain and meninges, oral pharynx, lung and bronchial tree,gastrointestinal tract, male and female reproductive tract, muscle,bone, skin, connective tissue, immune system, blood forming cells andbone marrow, liver and urinary tract, and special sensory organs such asthe eye. More specifically, human patients suffering from renal cellcarcinoma, pancreatic carcinoma, breast cancer, prostate cancer,colorectal cancer, malignant mesothelioma, multiple myeloma, ovariancancer, or melanoma may be treated with the VEGF trap as describedbelow.

Combination Therapies

In numerous embodiments, a VEGF fusion protein trap antagonist may beadministered in combination with one or more additional compounds ortherapies, including a second VEGF trap molecule. Combination therapyincludes administration of a single pharmaceutical dosage formulationwhich contains a VEGF trap and one or more additional agents; as well asadministration of a VEGF trap and one or more additional agent(s) in itsown separate pharmaceutical dosage formulation. For example, a VEGF trapand a cytotoxic agent, a chemotherapeutic agent or a growth inhibitoryagent can be administered to the patient together in a single dosagecomposition such as a combined formulation, or each agent can beadministered in a separate dosage formulation. Where separate dosageformulations are used, the VEGF-specific fusion protein of the inventionand one or more additional agents can be administered concurrently, orat separately staggered times, i.e., sequentially.

The term “cytotoxic agent” as used herein refers to a substance thatinhibits or prevents the function of cells and/or causes destruction ofcells. The term is intended to include radioactive isotopes (e.g. I¹³¹,I¹²⁵, Y⁹⁰ and Re¹⁸⁶), chemotherapeutic agents, and toxins such asenzymatically active toxins of bacterial, fungal, plant or animalorigin, or fragments thereof.

A “chemotherapeutic agent” is a chemical compound useful in thetreatment of cancer. Examples of chemotherapeutic agents includealkylating agents such as thiotepa and cyclosphosphamide (Cytoxan®);alkyl sulfonates such as busulfan, improsulfan and piposulfan;aziridines such as benzodopa, carboquone, meturedopa, and uredopa;ethylenimines and methylamelamines including altretamine,triethylenemelamine, trietylenephosphoramide, triethylenethiophosphaoramide and trimethylolomelamine; nitrogen mustards such aschlorambucil, chlornaphazine, cholophosphamide, estramustine,ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride,melphalan, novembichin, phenesterine, prednimustine, trofosfamide,uracil mustard; nitrosureas such as carmustine, chlorozotocin,fotemustine, lomustine, nimustine, ranimustine; antibiotics such asaclacinomysins, actinomycin, authramycin, azaserine, bleomycins,cactinomycin, calicheamicin, carabicin, caminomycin, carzinophilin,chromomycins, dactinomycin, daunorubicin, detorubicin,6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin,idarubicin, marcellomycin, mitomycins, mycophenolic acid, nogalamycin,olivomycins, peplomycin, potfiromycin, puromycin, quelamycin,rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex,zinostatin, zorubicin; anti-metabolites such as methotrexate and5-fluorouracil (5-FU); folic acid analogues such as denopterin,methotrexate, pteropterin, trimetrexate; purine analogs such asfludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidineanalogs such as ancitabine, azacitidine, 6-azauridine, carmofur,cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine;androgens such as calusterone, dromostanolone propionate, epitiostanol,mepitiostane, testolactone; anti-adrenals such as aminoglutethimide,mitotane, trilostane; folic acid replenisher such as frolinic acid;aceglatone; aldophosphamide glycoside; aminolevulinic acid; amsacrine;bestrabucil; bisantrene; edatraxate; defofamine; demecolcine;diaziquone; elformithine; elliptinium acetate; etoglucid; galliumnitrate; hydroxyurea; lentinan; lonidamine; mitoguazone; mitoxantrone;mopidamol; nitracrine; pentostatin; phenamet; pirarubicin; podophyllinicacid; 2-ethylhydrazide; procarbazine; PSK®; razoxane; sizofuran;spirogermanium; tenuazonic acid; triaziquone;2,2′,2″-trichlorotriethylamine; urethan; vindesine; dacarbazine;mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine;arabinoside (“Ara-C”); cyclophosphamide; thiotepa; taxanes, e.g.paclitaxel (Taxol®, Bristol-Myers Squibb Oncology, Princeton, N.J.) anddocetaxel (Taxotere®; Aventis Antony, France); chlorambucil;gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinumanalogs such as cisplatin and carboplatin; vinblastine; platinum;etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine;vinorelbine; navelbine; novantrone; teniposide; daunomycin; aminopterin;xeloda; ibandronate; CPT-11; topoisomerase inhibitor RFS 2000;difluoromethylornithine (DMFO); retinoic acid; esperamicins;capecitabine; and pharmaceutically acceptable salts, acids orderivatives of any of the above. Also included in this definition areanti-hormonal agents that act to regulate or inhibit hormone action ontumors such as anti-estrogens including for example tamoxifen,raloxifene, aromatase inhibiting 4(5)-imidazoles, 4-hydroxytamoxifen,trioxifene, keoxifene, LY 117018, onapristone, and toremifene(Fareston); and anti-androgens such as flutamide, nilutamide,bicalutamide, leuprolide, and goserelin; and pharmaceutically acceptablesalts, acids or derivatives of any of the above.

A “growth inhibitory agent” when used herein refers to a compound orcomposition which inhibits growth of a cell, especially a cancer celleither in vitro or in vivo. Examples of growth inhibitory agents includeagents that block cell cycle progression (at a place other than Sphase), such as agents that induce G1 arrest and M-phase arrest.Classical M-phase blockers include the vincas (vincristine andvinblastine), Taxol®, and topo II inhibitors such as doxorubicin,epirubicin, daunorubicin, etoposide, and bleomycin. Those agents thatarrest G1 also spill over into S-phase arrest, for example, DNAalkylating agents such as tamoxifen, prednisone, dacarbazine,mechlorethamine, cisplatin, methotrexate, 5-fluorouracil, and ara-C.

Pharmaceutical Compositions

Pharmaceutical compositions useful in the practice of the method of theinvention include a therapeutically effective amount of an active agent,and a pharmaceutically acceptable carrier. The term “pharmaceuticallyacceptable” means approved by a regulatory agency of the Federal or astate government or listed in the U.S. Pharmacopeia or other generallyrecognized pharmacopeia for use in animals, and more particularly, inhumans. The term “carrier” refers to a diluent, adjuvant, excipient, orvehicle with which the therapeutic is administered. Such pharmaceuticalcarriers can be sterile liquids, such as water and oils, including thoseof petroleum, animal, vegetable or synthetic origin, such as peanut oil,soybean oil, mineral oil, sesame oil and the like. Suitablepharmaceutical excipients include starch, glucose, lactose, sucrose,gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerolmonostearate, talc, sodium chloride, dried skim milk, glycerol,propylene, glycol, water, ethanol and the like. The composition, ifdesired, can also contain minor amounts of wetting or emulsifyingagents, or pH buffering agents. These compositions can take the form ofsolutions, suspensions, emulsion, tablets, pills, capsules, powders,sustained-release formulations and the like. The composition can beformulated as a suppository, with traditional binders and carriers suchas triglycerides. Oral formulation can include standard carriers such aspharmaceutical grades of mannitol, lactose, starch, magnesium stearate,sodium saccharine, cellulose, magnesium carbonate, etc. Examples ofsuitable pharmaceutical carriers are described in “Remington'sPharmaceutical Sciences” by E. W. Martin.

In a preferred embodiment, the composition is formulated in accordancewith routine procedures as a pharmaceutical composition adapted forintravenous, subcutaneous, or intramuscular administration to humanbeings. Where necessary, the composition may also include a solubilizingagent and a local anesthetic such as lidocaine to ease pain at the siteof the injection. Where the composition is to be administered byinfusion, it can be dispensed with an infusion bottle containing sterilepharmaceutical grade water or saline. Where the composition isadministered by injection, an ampoule of sterile water for injection orsaline can be provided so that the ingredients may be mixed prior toadministration.

The active agents of the invention can be formulated as neutral or saltforms. Pharmaceutically acceptable salts include those formed with freeamino groups such as those derived from hydrochloric, phosphoric,acetic, oxalic, tartaric acids, etc., and those formed with freecarboxyl groups such as those derived from sodium, potassium, ammonium,calcium, ferric hydroxides, isopropylamine, triethylamine, 2-ethylaminoethanol, histidine, procaine, etc.

Other features of the invention will become apparent in the course ofthe following descriptions of exemplary embodiments which are given forillustration of the invention and are not intended to be limitingthereof.

EXAMPLES

The following example is put forth so as to provide those of ordinaryskill in the art with a complete disclosure and description of how tomake and use the methods and compositions of the invention, and are notintended to limit the scope of what the inventors regard as theirinvention. Efforts have been made to ensure accuracy with respect tonumbers used (e.g., amounts, temperature, etc.) but some experimentalerrors and deviations should be accounted for. Unless indicatedotherwise, parts are parts by weight, molecular weight is averagemolecular weight, temperature is in degrees Centigrade, and pressure isat or near atmospheric.

Example 1 Pharmacokinetics and Safety of the VEGF Fusion Protein TrapAntagonist in Primates

Preclinical toxicology studies were conducted with the VEGF trap (SEQ IDNO:2) in primates and rodents. Four- and thirteen week toxicologystudies in cynomolgus monkeys showed that the VEGF trap was welltolerated when administered subcutaneously three times per week at dosesof 1.5, 5, and 15 mg/kg (four week study), or twice a week at 1.5, 5, 15or 30 mg/kg in the thirteen-week study. The VEGF trap was not highlyimmunogenic after four weeks in monkeys; only one mid-dose animaldeveloped low titer antibodies.

Example 2 Treatment of Solid Tumors or Non-Hodgkin's Lymphoma

Patients with refractory solid tumors or non-Hodgkin's lymphomareceiving no concurrent treatment for their cancer are treated with theVEGF trap as follows. The dose levels range from 0.3 mg/kg to 30 mg/kggiven subcutaneously. Each patient receives a single initial dose of theVEGF trap followed by four weeks of observation and pharmacokineticblood sampling. Beginning in the fifth week of the study, patientsreceive a series of 6 weekly injections at the assigned dose level.Plasma levels of the VEGF trap, and VEGF, both free and bound togetheras a complex, are monitored. Tumor burden is assessed at the beginningand end of the weekly dosing period and periodically during treatment;patients with stable disease, partial or complete responses may continuedosing for up to an additional 6 months in a continuation study. Athigher dose levels where efficacy might be anticipated, patients undergoDynamic Contrast Enhanced MRI scans to assess effects of VEGF trapadministration on tumor perfusion.

1. A method of treating a human patient suffering from renal cellcarcinoma, comprising administering an effective amount of a vascularendothelial growth factor (VEGF) antagonist VEGFR1R2-FcΔC1 (a) to thehuman patient, the method comprising: (a) administering to the patientan initial dose of at least approximately 0.5 to 10 mg/kg of the VEGFantagonist; and (b) administering to the patient a plurality ofsubsequent doses of the VEGF antagonist in an amount that isapproximately the same or less of the initial dose, wherein thesubsequent doses are separated in time from each other by at least oneday.
 2. The method of claim 1, wherein the initial dose is selected fromthe group consisting of approximately 1 mg/kg, 2 mg/kg, 3 mg/kg, 4mg/kg, 5 mg/kg and 6 mg/kg.
 3. The method of claim 1, wherein subsequentdoses are separated in time from each other by at least two weeks. 4.The method of claim 1, wherein the VEGF antagonist is VEGFR1R2-FcΔC1comprising the amino acid sequence of SEQ ID NO:
 2. 5. The method ofclaim 1, wherein administration is intravenous.
 6. The method of claim1, further comprising administering a second therapeutic agent.
 7. Themethod of claim 6, wherein the second therapeutic agent is a cytotoxicagent, a chemotherapeutic agent or a growth inhibitory agent.